In 1942 Albright and his associates described the features of a new clinical syndrome "pseudohypoparathyroidism" (PHP). Patients with this disorder show characteristic constitutional features (Albright's hereditary osteodystrophy - AHO) and do not respond to exogenous parathyroid hormone (PTH). In PHP, UcAMP (urinary cyclic AMP) does not increase normally in response to PTH administration. This indicates that there is a defective hormone receptor-adenylate cyclase complex in this disorder. We have shown that many patients with PHP+AHO (PHP Ia) show an approximately 50% reduction in activity of Gs (the stimulatory guanine nucleotide binding protein associated with adenylate cyclase) in membranes from multiple tissues. Gs deficiency presumably accounts for resistance to multiple hormones in such patients. Using cloned human cDNA probes for the alpha subunit of Gs, we have shown that steady state mRNA levels from fibroblasts of subjects with PHP Ia are reduced by approximately 50% compared with normals. We have now succeeded in defining the genetic abnormality responsible for Gs deficiency. Using the polymerase chain reaction to amplify genomic fragments encompassing each exon of the Gs-alpha gene, and comparing such fragments from normal and affected subjects on denaturing gradient gel electrophoresis, we were able to identify fragments with abnormal mobility. By direct DNA sequencing such fragments contained mutations that would explain reduction in mRNA in affected subjects.